Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
  • A61K31/42—Oxazoles
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
  • A61P35/02—Antineoplastic agents specific for leukemia

Definitions

• This invention relates to the use of 3-(7-oxo-1-aza-4-oxabicyclo[3.2.0]hept-3-yl) alanine derivatives as antitumor agents.
• G0069A JP 61-212587
• G0069A is a chemicalyl unstable isolation process and required very complex and special techniques. This should be done in the dark at low temperatures.
• the synthetic approach also seemed to be an extremely difficult multi-step process because they have 5-asymmetric carbon centres and dipeptide side chain. Therefore, it is necessary to get compounds which are relatively easy to synthesize, have shorter chains than G0069A, chemically stable and have stronger antitumor activity.
• the present invention relates to an antitumor composition
• an antitumor composition comprising of an effective amount of the 3-(7-oxo-1-aza-4-oxabicyclo[3.2.0]hept-3-yl)alanine derivative represented by the formula (I) or a pharmaceutically acceptable salt thereof ##STR2## and a pharmaceutically acceptable carrier, wherein R is:
• R 2 is C 1 -C 3 alkyl group which may be substituted with 1-3 aryl groups;
• R 1 is:
• Example of C 1 -C 3 alkyl group as substituent in R 1 and R 2 are methyl, ethyl, propyl or isopropyl.
• R in general formula (I) is selected from hydrogen, methoxycarbonyl, ethoxycarbonyl or benzyloxycarbonyl
• R 1 is selected from hydrogen, methyl, benzyl, diphenylmethyl or triphenylmethyl.
• Examples of pharmaceutically acceptable salts are sodium, potassium, calcium, magnesium, hydrogen chloride, tartaric, fumaric, maleic, acetic, trifluoroacetic, citric, methanesufonic, trifluoromethanesulfonic, p-toluenesulfonic and so on.
• the present invention provides a method of treating tumors in mammalian animals which comprises of administering to mammalian animals having tumors with an effective amount of the derivative of formula (I).
• the present invention provides use of the derivative of formula (I) for the preparation of a pharmacological composition for treatment of tumors.
• the bicyclic nucleus carries two asymmetric carbon atoms at position 3 and 5 and can exist as 4-diastereoisomers.
• the preferred isomer is (3R,5S) and (3S,5R) or mixture of them for superior toxicity against different malignant cells such as P388, KB, NUGC4, WI38, L-1210, sarcoma 180 and colon 26.
• Such diastereoisomers and their mixtures are also included within the use of oxapenam derivatives as antitumor agents.
• the chain alanine at C 3 of bicyclic nucleus carries one asymmetric carbon atom having D and L isomers. Both of the isomers (D and L) are included within the use of oxapenam derivatives as antitumor agents.
• Antitumor activity of the compounds described above is expected against some solid cancers such as gastrointestinal tract, lung, breast, liver, uterus and leukemia and so on.
• the present invention relates to the use of oxapenam derivatives having excellent antitumor activity.
• the compounds of this invention are characterized by the general formula (I) ##STR3##
• the synthesis of the compound of general formula (I) was done by following the synthetic scheme as shown below using DL-allyl-glycine as a starting material. ##STR4##
• the R and R 1 are the same as defined above.
• the R 3 is substituted sulfonyl group such as methanesulfonyl, trifluoromethanesulfonyl, bezenesulfonyl, 4-chlorobenzenesulfonyl, p-toluenesulfonyl, and so on.
• X is halogen atom such as fluorine, bromine, chlorine or iodine.
• M is metal such as sodium, potassium, lithium, and so on.
• the reactants are reacted together with solvent at elevated or low temperatures for sufficient time to allow the reaction to proceed to completion.
• the reaction conditions depend upon the nature and reactivity of the reactants.
• they are selected from triethylamine, pyridine, 4-diaminopyridine, diisopropylethylamine, 2,6-colidine, imidazole, piperidine, piperadine, pyrrolidine, morpholine, 1,8-diazabicyclo[5.4.0.]undec-7-ene, 1,5-diazabicyclo-[4.3.0]non-5-ene, sodium carbonate, potassium carbonate, lithium carbonate, cesium carbonate sodium hydrogen carbonate, potassium hydrogen carbonate, lithium hydrogen carbonate, cesium hydrogen carbonate and so on.
• the solvents of choice for the reactions are non reactive solvents such as acetonitrile, tetrahydrofuran, ethanol, methanol, benzene, chloroform, ethyl acetate, acetone, methylene chloride, water, dimethylformamide, dimethylsulfoxide, hexamethylphosphoric triamide, or the like. Solvent mixtures may also be utilized.
• Reaction temperatures would generally range from between -20° C. to 140° C.
• the preferred molar ratio of the reactants are 1:1 to 5.0.
• the reaction time range is from 0.5 to 72 h, depending on the reactants.
• the oxidizing agents are used for dihydroxylation of double bonds and are selected from either osmium tetroxide, potassium osmate, potassium permanganate, t-butyl hydroperoxide, hydrogen peroxide, AD mix- ⁇ , or AD mix- ⁇ .
• the AD mix- ⁇ and ⁇ may be used to prepare chiral diol 3 (J. Org. Chem. 57, 2768 (1992); Tetrahedron Lett. 34, 2267 (1993)).
• the deprotection of N and O protective group is carried out either by hydrogenation or by hydrolysis with mineral acids like hydrochloric acid in solvent like methanol, ethanol, propanol, ethyl acetate.
• the hydrogenation reaction is usually carried out in the presence of a metal catalyst such as Pd, Pt, Rh under normal pressure to high pressure of hydrogen.
• the compound of the invention when used as an agent for treating malignant tumors of mammals including humans, may take pharmaceutical dosage forms including parenteral preparations such as injections, suppositories, aerosols and the like and oral preparations such as tablets, coated tablets, powders, granules, capsules, liquids and the like. Injections are generally preferred.
• parenteral preparations such as injections, suppositories, aerosols and the like
• oral preparations such as tablets, coated tablets, powders, granules, capsules, liquids and the like. Injections are generally preferred.
• the above preparations are formulated in a manner known in the art.
• an excipient for the formulation of solid preparations for oral administration an excipient, and if desired, a binder, disintegrator, lubricant, coloring agent, corrigent, flavor, etc. are added to the compound of the invention, and then tablets, coated tablets, granules, powders, capsules or the like are prepared in a conventional manner.
• a pH adjusting agent, buffer, stabilizer, isotonic agent, local anesthetic or the like is added to the active ingredient of the invention, and injections for subcutaneous, intramuscular or intravenous administration can be prepared in a conventional manner.
• a base for the formulation of suppositories, a base, and if desired, a surfactant are added to the active ingredient of the invention, and the suppositories are prepared in a conventional manner.
• excipients useful for the solid preparations for oral administration are those generally used in the art and useful examples are excipients such as lactose, sucrose, sodium chloride, starches, calcium carbonate, kaolin, crystalline cellulose, methyl cellulose, glycerin, sodium alginate, gum arabic and the like, binders such as polyvinyl alcohol, polyvinyl ether, polyvinyl pyrrolidone, ethyl cellulose, gum arabic, schellac, sucrose, water, ethanol, propanol, carboxymethylcellulose, potassium phosphate and the like, lubricants such as magnesium stearate, talc and the like, and further include additives such as usual known coloring agents, disintegrators and the like.
• excipients such as lactose, sucrose, sodium chloride, starches, calcium carbonate, kaolin, crystalline cellulose, methyl cellulose, glycerin, sodium alginate, gum arabic and the like
• binders
• bases useful for the formulation of suppositories are oleaginous bases such as cacao butter, polyethylene glycol, lanolin, fatty acid triglycerides, Witepsol (trademark, Dynamite Nobel Co., Ltd.) and the like.
• Liquid preparations may be in the form of aqueous or oleaginous suspension, solution, syrup, elixir and the like, which can be prepared by a conventional way using usual additives.
• the amount of the compound (I) of the invention to be incorporated into the pharmaceutical composition of the invention varies with the dosage form, solubility and chemical properties of the compound, administration route, administration scheme and the like.
• the amount is about 1 to 25 w/w % in the case of oral preparations, and about 0.1 to about 5 w/w % in the case of injections which are parenteral preparations.
• the dosage of the compound (I) of the invention is suitably determined depending on the individual cases taking symptom, age and sex of the subject and the like into consideration.
• the dosage in the case of oral administration is about 50 to about 1000 mg per day for an adult in 2 to 4 divided doses
• the dosage in the case of injection, for example, by intravenous administration is 2 ml (about 1 to about 50 mg) which is administered once a day for an adult wherein the injection may be diluted with physiological saline or glucose injection liquid if so desired, and slowly administered over at least 5 minutes.
• the dosage in the case of suppositories is about 1 to about 500 mg which is administered once or twice a day at an interval of 6 to 12 hours wherein the suppositories are administered by insertion into the rectum.
• N-methylmorpholine N-oxide (12.6 ml) and osmium tetraoxide (4% wt soln. in water) (5 ml) was added to a solution of (DL)-N-(benzyloxycarbonyl)-allylglycine diphenylmethyl ester (25.57 g, 61.5 mmol) in water (30 ml) --acetone (240 ml).
• the mixture was stirred overnight and quenched with saturated sodium bisulfite solution (50 ml). After stirring for 10 minutes, the mixture was extracted with ethyl acetate (3 ⁇ 150 ml), washed with brine, dried (MgSO 4 ) and the solvent was removed in vacuo. Purification by a silica gel column chromamography using hexane-ethyl acetate (1:4) as The eluant gave 3 (20.74 g, 70%) as oil.
• Triethylamine (8.19 g, 81 mmol) and palladium (II) acetate (1.82 g, 8.1 mmol) was added to a solution of 4-acetoxyazetidinone (10.52 g, 81 mmol) and the alcohol 4 (21.83 g, 41 mmol) in benzene (500 ml).
• the mixture was stirred at room temperature under nitrogen atmosphere for 20 hr and filtered through a pad of celite.
• the celite was washed with ethyl acetate (300 ml) and the combined organic layer was washed with brine (3 ⁇ 150 ml), dried (MgSO 4 ) and the solvent was removed in vacuo.
• Purification by silica gel column chromatography using hexane-ethyl acetate (1:1) as eluant gave ⁇ (16.76 g 55%) as a foam.
• Tetrabutylammonium fluoride (1M solution in THF) 40 ml, 46.2 mmol
• glacial acetic acid 5 ml
• the mixture was stirred at room temperature for 4 h.
• the solvent was concentrated and the residue was loaded onto a silica gel column. Elution with hexane-ethyl acetate (1:1) removed impurities.
• the desired alcohol 6 (10.1 g, 63%) was obtained as foam after eluting with ethyl acetate-acetone (4:1).
• p-Toluenesulfonyl chloride 14.96 g, 26 mmol was added to a solution of the alcohol 6 (9.0 g, 17.2 mmol) in pyridine (42 ml) cooled to -10° C. The resulting mixture was stirred for 4 h and poured onto a cold 2N HCl (600 ml) solution. The mixture was extracted with ethyl acetate (3 ⁇ 200 ml) and the ethyl acetate portion was washed with water (100 ml), brine, dried (MgSO 4 ) and the solvent was removed in vacuo. Purification by silica gel column chromatography using hexane-ethyl acetate (1:1) as the eluant gave 7 (9.92 g, 85%) as white foam.
• Lithium bromide was added to a solution of tosylate 7 (1.9 9, 2.82 mmol) in hexamethylphospholic triamide (HMPA) (20 ml) and the mixture was heated at 60° C. under nitrogen atmosphere for 3 h. The solution was poured into cold water (250 ml) and extracted with ethyl acetate (3 ⁇ 150 ml). The ethyl acetate portion was washed with water (3 ⁇ 100 ml), brine, dried (MgSO 4 ) and the solvent was removed in vacuo. Purification by silica gel column chromatography using hexane-ethyl acetate as the eluant gave 9 (1.02 9, 62%) as a white foam.
• HMPA hexamethylphospholic triamide
• Triethylamine (0.52 ml, 3.71 mmol) and palladium (II) acetate (0.083 g, 0.37 mmol) was added to a stirred solution of 4-acetoxy azetidinone (0.48 g, 3.71 mmol) and bromohydrin 8 (0.95 g, 1.85 mmol) in benzene (50 ml).
• the mixture was stirred for 20 h at room temperature under nitrogen atmosphere and filtered through a pad of celite.
• the celite was washed with ethyl acetate (100 ml) and the combined organic layer was washed with water (40 ml), brine, dried (MgSO 4 ) and the solvent was removed in vacuo.
• Purification by silica gel column chromatography using hexane-ethyl acetate (1:1) as the eluant gave 9 (0.22 g, 30%) as a foam.
• KB cells were cultivated in Eagles minimum essential medium supplemented with 10% calf serum and incubated at 37° C. in a humidified 5% CO 2 atmosphere to prepare a cell stock. Cells were counted using a neubauer hemocytometer and seeded in 96 well plates at 100 ⁇ l of 3 ⁇ 10 4 cells/ml and cultured for one day. Test compounds were diluted and 100 ⁇ l of the solution was added in triplicate wells to give final concentration of 10, 5, 1, 0.5, 0.1, 0.05 and 0.01 ⁇ g/ml. Control wells were identical except that test compound was absent. These were cultured for three days. Then the cells were fixed with addition of 20 ⁇ l of 25% glutaraldehyde for 15 minutes, washed with water and dried.
• L1210 cells were cultivated in RPMI 1640 medium supplemented with 10% fetal calf serum and 50 ⁇ l of 2-mercaptoethanol at 37° C. in humidified 5% CO 2 atmosphere no prepare a cell stock. Cells were counted using neubauer hemocytometer and seed in 96 well plates at 100 ⁇ l of 0.5 ⁇ 10 4 cells per ml. The test compounds were diluted and 100 ⁇ l of the solution was added in triplicate wells to give the final concentration of 10, 5, 1, 0.5, 0.1, 0.05 and 0.01 ⁇ g/ml. Control wells were identical except :hat the test compound was absent. These were cultured for three days. Results were assayed using the microculture tetrazolium assay briefly.
• TD 50 values were calculated from linear depression ines of the log-logit plot.
• the compounds of general formula (I) were tested in vivo against Sarcoma 180 xenografted tumor to mice as illustrated herein after.
• Sarcoma 180 5 ⁇ 10 6 cells were inoculated by S.C. to male ICR mice (6 weeks old) on day 0. Drugs were administered on days 1,5 and 9. Mice were killed and tumor weight was measured on day 12 after transplantation. The percentage inhibition of tumor growth was calculated from the mean tumor weight of the treated group compared with that of the control group. Number of mice used in each group was between 6 to 10. The percentage inhibition of tumor Sarcoma 180 group by compound of formula (I) are summarized in Table 2.

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• 1994
  • 1994-01-07 GB GB9400239A patent/GB9400239D0/en active Pending
• 1995
  • 1995-01-06 JP JP7518374A patent/JPH08507551A/ja active Pending
  • 1995-01-06 AU AU13249/95A patent/AU688441B2/en not_active Ceased
  • 1995-01-06 CA CA002157603A patent/CA2157603A1/fr not_active Abandoned
  • 1995-01-06 WO PCT/GB1995/000023 patent/WO1995018611A1/fr not_active Ceased
  • 1995-01-06 US US08/513,803 patent/US5652234A/en not_active Expired - Fee Related
  • 1995-01-06 EP EP95904657A patent/EP0690714A1/fr not_active Ceased
  • 1995-01-09 ZA ZA95107A patent/ZA95107B/xx unknown

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